CN102150741B - Method for preparing lactalbumin hydrolysate by utilizing yak milk - Google Patents
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Abstract
The invention provides a method for preparing lactalbumin hydrolysate by utilizing yak milk, and belongs to the field of milk product processing. The method for preparing the lactalbumin hydrolysate finished product comprises the following steps: degreasing the yak milk, preparing rennet casein, baking and crushing; hydrolyzing, inactivating enzyme, purifying, concentrating, drying and carrying out other steps. The method for preparing the lactalbumin hydrolysate has the advantages of short time as well as mild reacting condition, complete hydrolysis and high hydrolyzing rate by using bienzyme hydrolysis compared with acid and alkali hydrolysis.
Description
Technical field
The present invention relates to the processing of dairy product, relate in particular to a kind of working method of yak whey solution milk-protein.
Background technology
Have only U.S. Gibco and Hyclone two companies to utilize cow's milk to produce lactoalbumin hydrolysate abroad, product is found a good sale in countries such as Europe, the United States, Asia.The production of domestic lactoalbumin hydrolysate is blank, and nearly all scientific research and production user be the dependence on import product, and imported product is expensive, has increased operation cost greatly.And traditional technology hydrolysis cow's milk uses strong acid and highly basic hydrolysis casein food grade, and bronsted lowry acids and bases bronsted lowry all is chemical substance on the one hand, causes certain environmental pollution, on the other hand, strong acid and highly basic effect are strong, and hydrolysis degree is difficult for grasping, destroy amino acid easily, percent hydrolysis is low, the mortality height.
Summary of the invention
The technical problem to be solved in the present invention is to overcome the defective that existing yak breast utilizes, and the working method of the workable yak whey solution milk-protein of a kind of economic benefit height, method is provided.
In order to solve the problems of the technologies described above, the invention provides following technical scheme:
A kind of method of utilizing the yak breast to prepare lactoalbumin hydrolysate comprises the steps:
(1), degreasing: be raw material with bright yak breast, the yak breast is collected in the degreasing machine degreasing;
(2), preparation rennin casein food grade: the yak Ruzhong after step (1) degreasing adds calcium chloride solution, transfers PH5.5～6.5, is positioned over 35～45 ℃ of water-baths then, adds calf chymosin solution, stir make the yak curdling solid the rennin casein food grade;
(3), with rennin casein food grade oven dry in (2), pulverizing;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pH9.0～11.0 by 4-8% concentration, heating is dissolved the rennin casein food grade fully, pH value of solution is maintained 7.5～8.5, in E:S=1:(6～14) ratio adding trypsinase, each aminoacids content of high-performance liquid chromatogram determination is used in hydrolysis, calculate degree of hydrolysis, degree of hydrolysis reaches 70%.Again pH value of solution is transferred to 8.5～9.5, in E:S=1:(40～80) ratio adding prozyme, hydrolysis; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 98%; Described prozyme refers to that Chymotrypsin, stomach en-, trypsinase are with mass ratio 1:(1～3): mix (0.5～1.5);
(5), inactivator: the hydrolyzed solution of step (4) is boiled inactivator;
(6), filter: with enzyme and the unhydrolysed albumen of the feed liquid filtering sex change of step (5);
(7), purifying: ultrafiltration, collection filtered solution;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 6.8～7.2;
(9), drying: 110～150 ℃ of following spraying dryings, moisture gets the lactoalbumin hydrolysate finished product less than 5% with concentrated solution in the step (8).Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis is greater than 90%, and it is qualified to be;
Preferably, described calf chymosin is at the weightmeasurement ratio 0.0005～0.0015% in yak Ruzhong, and described tryptic add-on standard is E:S=1:10, and prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:2:1.
The method time spent of lactoalbumin hydrolysate of the present invention is short, uses double-enzyme hydrolysis acid, basic hydrolysis, the reaction conditions gentleness, and hydrolysis is thorough, the percent hydrolysis height.Main purpose of the present invention is to take full advantage of local yak breast resource, existing cow's milk and casein food grade product are carried out scientific and technical innovation, industrial upgrading, use the double-enzyme hydrolysis casein, lactoalbumin hydrolysate and the cell substratum product of the calf chymosin casein food grade development high added value through also using biological level.The lactoalbumin hydrolysate that is got by rennin casein food grade institute's enzymolysis of biological level, the biologically active peptides that obtains behind the enzymolysis is the added value height not only, and security is good, and technology is simple, and carries out industrialization easily.
Embodiment
Below in conjunction with accompanying drawing the preferred embodiments of the present invention are described, should be appreciated that preferred embodiment described herein only is used for description and interpretation the present invention, and be not used in restriction the present invention.
Embodiment 1
A kind of method of utilizing the yak breast to prepare lactoalbumin hydrolysate, step is as follows:
(1), degreasing: be raw material with bright yak breast, use the degreasing machine degreasing, rotating speed 3000～4000r/min, centrifugation time 25～35min, the filtering precipitation is collected supernatant liquor;
(2), preparation rennin casein food grade: it is the calcium chloride of 0.01～0.02mol/L that the yak Ruzhong after step (1) degreasing adds concentration, transfer PH5.5～6, be positioned over 39～41 ℃ of water bath heat preservation 25～35min then, described calf chymosin is that 0.001%(adds the 1g calf chymosin in the cow's milk of every 100000ml at the weightmeasurement ratio in yak Ruzhong), stir make the yak curdling solid the rennin casein food grade;
(3), rennin casein food grade in (2) is dried 4～6h in 60～70 ℃ of vacuum drying ovens, the pulverized particles degree is 200 orders;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pH10.0 by 6% concentration, 60～70 ℃ of heating are dissolved the rennin casein food grade fully, pH value of solution is maintained 7.8～8.0,55 ℃ of temperature add trypsinase, hydrolysis 5h in the E:S=1:10 ratio; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 70%.Again pH value of solution is transferred to 9.0～9.5,55 ℃ of temperature add prozyme in the E:S=1:60 ratio, keep 5h; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 98%; Prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:2:1;
(5), inactivator: the hydrolyzed solution of step (4) is boiled 10min, inactivator;
(6), filter: the feed liquid of step (5) with 3800r/min centrifuging 30min, is removed enzyme and the unhydrolysed albumen of sex change;
(7), purifying: use molecular weight cut-off to be the film bag ultrafiltration of 1KD, collect filtered solution, remove the polypeptide of the halfway macromolecule of hydrolysis;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 7.0～7.2;
(9), drying: with concentrated solution in the step (8) 120～140 ℃ of following spraying dryings, to moisture less than 5%, get the lactoalbumin hydrolysate finished product.
Products obtained therefrom uses each aminoacids content of high-performance liquid chromatogram determination, calculates degree of hydrolysis, degree of hydrolysis 95%.
Embodiment 2
A kind of method of utilizing the yak breast to prepare lactoalbumin hydrolysate, step is as follows:
(1), degreasing: be raw material with bright yak breast, use the degreasing machine degreasing, rotating speed 3000～4000r/min, centrifugation time 25～35min, the filtering precipitation is collected supernatant liquor;
(2), preparation rennin casein food grade: it is the calcium chloride of 0.01～0.02mol/L that the yak Ruzhong after step (1) degreasing adds concentration, transfer PH6.0～6.5, be positioned over 35～40 ℃ of water bath heat preservation 25～35min then, described calf chymosin is that 0.0005%(adds the 0.5g calf chymosin in the cow's milk of every 100000ml at the weightmeasurement ratio in yak Ruzhong), stir make the yak curdling solid the rennin casein food grade;
(3), rennin casein food grade in (2) is dried 4～6h in 60～70 ℃ of vacuum drying ovens, the pulverized particles degree is 200 orders;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pH9.0 by 4% concentration, 60～70 ℃ of heating are dissolved the rennin casein food grade fully, pH value of solution is maintained 7.5～8.0,55 ℃ of temperature add trypsinase, hydrolysis 5h in the E:S=1:6 ratio; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 70%.Again pH value of solution is transferred to 8.5～9.0,55 ℃ of temperature add prozyme in the E:S=1:40 ratio, keep 5h; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 96%; Prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:1:1.5;
(5), inactivator: the hydrolyzed solution of step (4) is boiled 10min, inactivator;
(6), filter: the feed liquid of step (5) with 3800r/min centrifuging 30min, is removed enzyme and the unhydrolysed albumen of sex change;
(7), purifying: use molecular weight cut-off to be the film bag ultrafiltration of 1KD, collect filtered solution, remove the polypeptide of the halfway macromolecule of hydrolysis;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 6.8～7.0;
(9), drying: with concentrated solution in the step (8) 110～130 ℃ of following spraying dryings, to moisture less than 5%, get the lactoalbumin hydrolysate finished product.
Products obtained therefrom uses each aminoacids content of high-performance liquid chromatogram determination, calculates degree of hydrolysis, degree of hydrolysis 94%.
Embodiment 3
A kind of method of utilizing the yak breast to prepare lactoalbumin hydrolysate, step is as follows:
(1), degreasing: be raw material with bright yak breast, use the degreasing machine degreasing, rotating speed 3000～4000r/min, centrifugation time 25～35min, the filtering precipitation is collected supernatant liquor;
(2), preparation rennin casein food grade: it is the calcium chloride of 0.01～0.02mol/L that the yak Ruzhong after step (1) degreasing adds concentration, transfer PH6.1～6.4, be positioned over 41～45 ℃ of water bath heat preservation 25～35min then, described calf chymosin is that 0.0015%(adds the 1.5g calf chymosin in the cow's milk of every 100000ml at the weightmeasurement ratio in yak Ruzhong), stir make the yak curdling solid the rennin casein food grade;
(3), rennin casein food grade in (2) is dried 4～6h in 60～70 ℃ of vacuum drying ovens, the pulverized particles degree is 180 orders;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pH11.0 by 6% concentration, 60～70 ℃ of heating are dissolved the rennin casein food grade fully, pH value of solution is maintained 8.1～8.5,55 ℃ of temperature add trypsinase, hydrolysis 5h in the E:S=1:14 ratio; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 70%.Again pH value of solution is transferred to 8.7～9.2,55 ℃ of temperature add prozyme in the E:S=1:80 ratio, keep 5h; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 99%; Prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:3:0.5;
(5), inactivator: the hydrolyzed solution of step (4) is boiled 10min, inactivator;
(6), filter: the feed liquid of step (5) with 3800r/min centrifuging 30min, is removed enzyme and the unhydrolysed albumen of sex change;
(7), purifying: use molecular weight cut-off to be the film bag ultrafiltration of 1KD, collect filtered solution, remove the polypeptide of the halfway macromolecule of hydrolysis;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 6.9～7.1;
(9), drying: with concentrated solution in the step (8) 130～150 ℃ of following spraying dryings, to moisture less than 5%, get the lactoalbumin hydrolysate finished product.
Products obtained therefrom uses each aminoacids content of high-performance liquid chromatogram determination, calculates degree of hydrolysis, degree of hydrolysis 96%.
It should be noted that at last: the above only is the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment the present invention is had been described in detail, for a person skilled in the art, it still can be made amendment to the technical scheme that aforementioned each embodiment puts down in writing, and perhaps part technical characterictic wherein is equal to replacement.Within the spirit and principles in the present invention all, any modification of doing, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (3)
1. a method of utilizing the yak breast to prepare lactoalbumin hydrolysate is characterized in that, may further comprise the steps:
(1), degreasing: be raw material with bright yak breast, use the degreasing machine degreasing, rotating speed 3000～4000r/min, centrifugation time 25～35min, the filtering precipitation is collected supernatant liquor;
(2), preparation rennin casein food grade: it is the calcium chloride of 0.01～0.02mol/L that the yak Ruzhong after step (1) degreasing adds concentration, transfer PH5.5～6, be positioned over 39～41 ℃ of water bath heat preservation 25～35min then, described calf chymosin is that 0.001%(adds the 1g calf chymosin in the cow's milk of every 100000ml at the weightmeasurement ratio in yak Ruzhong), stir make the yak curdling solid the rennin casein food grade;
(3), rennin casein food grade in (2) is dried 4～6h in 60～70 ℃ of vacuum drying ovens, the pulverized particles degree is 200 orders;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pHl0.0 by 6% concentration, 60～70 ℃ of heating are dissolved the rennin casein food grade fully, pH value of solution is maintained 7.8～8.0,55 ℃ of temperature add trypsinase, hydrolysis 5h in the E:S=1:10 ratio; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 70%, again pH value of solution is transferred to 9.0～9.5, and 55 ℃ of temperature add prozyme in the E:S=1:60 ratio, keep 5h; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 98%; Prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:2:1;
(5), inactivator: the hydrolyzed solution of step (4) is boiled l0min, inactivator;
(6), filter: the feed liquid of step (5) with 3800r/min centrifuging 30min, is removed enzyme and the unhydrolysed albumen of sex change;
(7), purifying: use molecular weight cut-off to be the film bag ultrafiltration of 1KD, collect filtered solution, remove the polypeptide of the halfway macromolecule of hydrolysis;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 7.0～7.2;
(9), drying: with concentrated solution in the step (8) 120～140 ℃ of following spraying dryings, to moisture less than 5%, get the lactoalbumin hydrolysate finished product.
2. a method of utilizing the yak breast to prepare lactoalbumin hydrolysate is characterized in that, may further comprise the steps:
(1), degreasing: be raw material with bright yak breast, use the degreasing machine degreasing, rotating speed 3000～4000r/min, centrifugation time 25～35min, the filtering precipitation is collected supernatant liquor;
(2), preparation rennin casein food grade: it is the calcium chloride of 0.01～0.02mol/L that the yak Ruzhong after step (1) degreasing adds concentration, transfer PH6.0～6.5, be positioned over 35～40 ℃ of water bath heat preservation 25～35min then, described calf chymosin is that 0.0005%(adds the 0.5g calf chymosin in the cow's milk of every l00000ml at the weightmeasurement ratio in yak Ruzhong), stir make the yak curdling solid the rennin casein food grade;
(3), rennin casein food grade in (2) is dried 4～6h in 60～70 ℃ of vacuum drying ovens, the pulverized particles degree is 200 orders;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pH9.0 by 4% concentration, 60～70 ℃ of heating are dissolved the rennin casein food grade fully, pH value of solution is maintained 7.5～8.0,55 ℃ of temperature add trypsinase, hydrolysis 5h in the E:S=1:6 ratio; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 70%, again pH value of solution is transferred to 8.5～9.0, and 55 ℃ of temperature add prozyme in the E:S=l:40 ratio, keep 5h; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 96%; Prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:1:1.5;
(5), inactivator: the hydrolyzed solution of step (4) is boiled l0min, inactivator;
(6), filter: the feed liquid of step (5) with 3800r/min centrifuging 30min, is removed enzyme and the unhydrolysed albumen of sex change;
(7), purifying: use and hold back the branch component and be the film bag ultrafiltration of 1KD, collect filtered solution, remove the polypeptide of the halfway macromolecule of hydrolysis;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 6.8～7.0;
(9), drying: with concentrated solution in the step (8) 110～130 ℃ of following spraying dryings, to moisture less than 5%, get the lactoalbumin hydrolysate finished product.
3. a method of utilizing the yak breast to prepare lactoalbumin hydrolysate is characterized in that, may further comprise the steps:
(1), degreasing: be raw material with bright yak breast, use the degreasing machine degreasing, rotating speed 3000～4000r/min, centrifugation time 25～35min, the filtering precipitation is collected supernatant liquor;
(2), preparation rennin casein food grade: it is the calcium chloride of 0.01～0.02mol/L that the yak Ruzhong after step (1) degreasing adds concentration, transfer PH6.1～6.4, be positioned over 41～45 ℃ of water bath heat preservation 25～35min then, described calf chymosin is that 0.0015%(adds the 1.5g calf chymosin in the cow's milk of every l00000ml at the weightmeasurement ratio in yak Ruzhong), stir make the yak curdling solid the rennin casein food grade;
(3), rennin casein food grade in (2) is dried 4～6h in 60～70 ℃ of vacuum drying ovens, the pulverized particles degree is 180 orders;
(4), take by weighing the rennin casein food grade of step (3), the rennin casein food grade is dissolved in the water of pHll.0 by 6% concentration, 60～70 ℃ of heating are dissolved the rennin casein food grade fully, pH value of solution is maintained 8.1～8.5,55 ℃ of temperature add trypsinase, hydrolysis 5h in the E:S=l:14 ratio; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 70%, again pH value of solution is transferred to 8.7～9.2, and 55 ℃ of temperature add prozyme in the E:S=1:80 ratio, keep 5h; Use each aminoacids content of high-performance liquid chromatogram determination, calculate degree of hydrolysis, degree of hydrolysis reaches 99%; Prozyme refers to that Chymotrypsin, stomach en-, trypsinase mix with mass ratio 1:3:0.5;
(5), inactivator: the hydrolyzed solution of step (4) is boiled l0min, inactivator;
(6), filter: the feed liquid of step (5) with 3800r/min centrifuging 30min, is removed enzyme and the unhydrolysed albumen of sex change;
(7), purifying: use molecular weight cut-off to be the film bag ultrafiltration of 1KD, collect filtered solution, remove the polypeptide of the halfway macromolecule of hydrolysis;
(8), concentrate: filtrate in the step (7) is concentrated, and regulating the pH value is 6.9～7.1;
(9), drying: with concentrated solution in the step (8) 130～150 ℃ of following spraying dryings, to moisture less than 5%, get the lactoalbumin hydrolysate finished product.
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| CN103798390B (en) * | 2014-01-23 | 2015-10-28 | 甘南倍益特生物科技有限公司 | A kind of yak milk casein nutrient powder and preparation method thereof being applicable to the elderly |
| CN105085650A (en) * | 2015-08-31 | 2015-11-25 | 苏州普罗达生物科技有限公司 | Small molecular milk casein polypeptide and preparing method |
| CN105111298A (en) * | 2015-08-31 | 2015-12-02 | 常州亚当生物技术有限公司 | Small molecule yak milk casein peptide and preparation method |
| CN105175502A (en) * | 2015-08-31 | 2015-12-23 | 常州亚当生物技术有限公司 | Small molecule yak milk whey protein polypeptide and preparation method thereof |
| CN112094880B (en) * | 2020-08-30 | 2022-03-29 | 中南林业科技大学 | Preparation method of yak milk residue polypeptide with antioxidant activity |
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| CN1260248C (en) * | 2004-09-03 | 2006-06-21 | 王秀英 | Active lactoprotein extracted from yak milk and its extraction method and use |
| CN100398660C (en) * | 2005-12-07 | 2008-07-02 | 广东省微生物研究所 | Method for producing compound bioactive peptide utilizing casein enzyme process |
| CN101041845B (en) * | 2006-12-09 | 2010-04-07 | 甘肃华羚干酪素有限公司 | Method for producing chymosin dry casein by using Qula as raw material |
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